计算机图像处理系统用于血管平滑肌细胞形态定量分析

采用计算机图像分析法，定量测定家兔培养主动脉血管平滑肌细胞的形态学参数及其与ＤＮＡ合成指标的相关关系，并在此基础上，从细胞形态学角度探讨了中药制剂川芎嗪对ＶＳＭＣ增殖的影响，以评价计算机图像分析技术在细胞学研究中的作用，建立一种研究ＶＳＭＣ增殖水平的新方法。     

Inhibitory effect and mechanism of chuanxiongzine on multiplication of VSMC

Objective: To study the inhibitory effect of chuanxiongzine on vascular smooth muscle cell (VSMC) proliferation and explore its molecular biology basis. Methods: we selected the VSMC cultured 4～8 generation from rat aorta thoracalis as research object.The objects were divided into four groups( Ⅰ )control group, ( Ⅱ )chuanxiongzine(50 μg/ml)group, ( Ⅲ )chuanxiongzine (100 μg/ml) group and( Ⅳ ) chuanxiongzine (200 μg/ml) group. The inhib itory effect of chuanxiongzine on VSMC proliferation was investigated by cell counting, MTT and 3H-TdR incorporation assay. In order to illuminate the molecular biology mechanism of chuanxiongzine inhibiting VSMCs proliferation, the expression of proliferating cell nuclear antigen (PCNA) and C-myc were detected.Results: Chuanxiongzine could inhibit the proliferation of VSMC significantly in a dose- and time-dependent manner, compared with control group (P ＜ 0.05). The expression of PCNA and c-myc were inhibited obviously and correlated with the concentration of chuanxiongzine (P ＜ 0.05). Conclusion: Chuanxiongzine may play a considerable role in VSMC proliferation process. The inhibitory effect of chuanxiongzine in a dose- and time-dependent manner can be realized via down regulating the expression of PCNA and c-myc. In this study, The great theoretical fundament about Chinese medicine, which is used to treat atherosclerosis (AS), has been obtained.     

降钙素基因相关肽抑制大鼠主动脉平滑肌细胞增殖

本工作用培养的大鼠主动脉平滑肌细胞，以细胞计数和＾３Ｈ－胸腺嘧啶掺入为指标，观察到降钙素基因相关肽呈剂量依赖地抑制ＶＳＭＣ增殖，１０＾－８和１０＾－７ｍｏｌ／ＬＣＧＲＰ可使＾３Ｈ－ＴｄＲ掺入量较对照组减少２６％（Ｐ＜０．０５）和３６％（Ｐ＜０．０１）并使细胞计数分别较对照组下降１７％（Ｐ＜０．０５）和３５％（Ｐ＜０．０１）。提示ＣＧＲＰ参与ＶＳＭＣ增殖的调节。     

Cytologic identification of clinically occult proliferative breast disease in women with a family history of breast cancer

A cytologic method for sampling the normal breast by fine-needle aspiration (FNA) was used to determine the frequency of clinically inapparent proliferative breast disease (PBD) in women with family histories of breast cancer. The authors attempted to obtain specimens from each quadrant of both breasts in 51 female first-degree relatives of breast cancer patients. The study group had no detectable masses by physical examination or mammography. Samples were prepared on membrane filters, Papanicolaou stained, and evaluated cytomorphologically. Three hundred seventy-eight of 408 (92.6%) possible quadrants were sampled; cellular material was obtained from 290 (76.7%) quadrants. PBD was identified in 20 of the 51 women (39.2%). When epithelium was obtained, nuclear area, perimeter, and diameter were measured with the use of computerized image analysis. Nuclei in samples containing atypical hyperplasia showed significant differences in these parameters when compared with cells from samples containing normal epithelium or benign hyperplasia. The authors' findings indicate that FNA sampling and computerized image analysis are useful in the detection and characterization of clinically inapparent PBD.     

一氧化氮在脂多糖抑制血管平滑肌细胞增殖中的作用

本研究采用MTT 法,3 H-TdR 掺入实验、分光光度计测量和免疫组织化学的方法,观察了脂多糖对离体培养SD 大鼠血管平滑肌细胞殖增、DNA 合成、一氧化氮产量和一氧化氮合酶表达的影响。结果发现,脂多糖可明显抑制血管平滑肌细胞的增殖和DNA 合成,推测该效应可能是通过iNOS-NO-cGMP 通路实现的。     

血管紧张素Ⅱ受体拮抗剂对高血压肾小动脉重建的影响

目的:研究血管紧张素II(AngII)受体拮抗剂对高血压肾小动脉重建的影响。方法:18只4周龄雄性大鼠分为:正常血压大鼠(WKY)组、自发性高血压大鼠(SHR)组、SHR口服losartan组,均饲养至16周。在肾组织切片上分别用光镜和电镜配合计算机图像分析法观测肾组织内小动脉的几何形态学指标和小动脉平滑肌及其间隙,离体肾脏灌流法测定最小肾血管阻力。结果:Losartan组的尾动脉收缩压、肾小动脉壁厚、壁面积、壁厚内径比和中层血管平滑肌细胞宽度以及最小肾血管阻力,均显著低于高血压对照组。结论:AngII受体拮抗剂losartan能预防SHR肾小动脉的重建。     

Ultrastructural morphometric analysis of papillary neoplasms: biological and diagnostic relevance

Ten papillary adenocarcinomas of thyroid origin (P-Thy), ten papillary adenocarcinomas of ovarian origin (P-Ov), and eight papillary neoplasms of non-thyroid/non-ovarian origin (P-Other) were morphometrically compared using 19 distinct quantitative nuclear and nucleolar parameters as a database for diagnosis. The selected cases consisted of 16 primary and 12 metastatic neoplasms. It was determined that the P-Thy group had a significantly smaller nucleolar area (NuA) and nucleolar perimeter (NuP), and smaller SDs of nuclear area (NA), NuA, and NuP compared with the P-Ov and P-Other groups (P less than .05). The P-Ov group had a significantly smaller SD of NA compared with the P-Other group (P less than .05). The P-Ov group exhibited the greatest variability among the papillary neoplasms. Linear regression analysis indicated that in the P-Thy group alone there was a significant correlation between mean nuclear form factor (4 pi A/P2) and mean NuA (r = -.82; P less than .01), and mean NP and mean NuA (r = +.77; P less than .01). Linear regression analysis also indicated that in the P-Ov group alone, there was a significant correlation between mean NA and mean NuA (r = +.75; P less than .02). Morphometric domains were established using statistically significant sets of variables that distinguished between the groups. The application of three-dimensional computerized cluster analysis techniques indicated that the P-Thy group consistently had the smallest morphometric domains. It was concluded that ultrastructural morphometric analysis of papillary neoplasms has diagnostic potential and reveals interesting biological relationships among distinct nuclear features in the different groups of neoplasms.     

Parotid gland tumors: Correlation between routine cytology and cytomorphometry by digital image analysis using conventional and newly introduced cytomorphometric parameters

The objective of this study was to compare qualitative cytomorphology and morphometric characteristics of parotid gland tumor cells, with the aid of a computer‐assisted system of image analysis. Routine qualitative cytologic and quantitative morphometric results from 64 parotid gland tumors were compared. Ultrasound (US)‐guided fine‐needle aspiration (FNA) specimens were taken from 54 patients. Eleven conventionally used morphometric parameters were studied: area, perimeter, convex area, convexity, maximal and minimal radius, length, breadth, form factor (FF), elongation factor, and nuclear‐ cytoplasmatic (N/C) ratio. Two newly introduced nuclear form factors were also measured: area symmetry factor and perimeter symmetry factor. The following nuclear morphometric parameters were significantly different between malignant and benign tumors: area, perimeter, convex area, convexity, maximal and minimal radius, length, breadth, FF, elongation factor, area symmetry factor, and perimeter symmetry factor. Comparing the cutoff values and receiver operating characteristic (ROC) curves the following nuclear morphometric parameters were found most useful in separating benign from malignant tumors: area, perimeter, convex area, maximal radius, length, and FF. The following whole cell morphometric parameters were significantly different between malignant and benign tumors: minimal and maximal radius, convexity, breadth, FF, and elongation factor. N/C ratio was significantly higher in malignant tumors. The quantitative morphometric analysis is a useful tool in the cytological differentiation between benign and malignant parotid gland tumors. Computerized image analysis may add to morphological evaluation by turning qualitative data into quantitative values. Diagn. Cytopathol. 2013;41:776–784. © 2013 Wiley Periodicals, Inc.     

Morphometric distinction of signet-ring cell adenocarcinoma cells from foamy macrophages in gastric endoscopic biopsies

AIMS : To distinguish signet-ring cancer cells from foamy macrophages in the small gastric endoscopic biopsies using objective morphometric measurements of nuclei. MATERIAL AND METHODS : Using computerized image analysis, the mean nuclear area, length, breadth, perimeter and roundness were analyzed in histological sections of ten gastric endoscopic biopsies with signet-ring cell adenocarcinoma and four benign lesions with numerous foamy macrophages. RESULTS : Nuclei of signet-ring cell adenocarcinoma were significantly bigger than nuclei of foamy macrophages. Mean nuclear area (34.25 μm2 for carcinoma cells vs. 25.41 μm2 for macrophages) and mean nuclear breadth (5.82 μm vs. 4.99 μm, respectively) differed significantly (P < 0.05), whereas the remaining parameters did not. CONCLUSION : Nuclear morphometry can distinguish foamy macrophages from signet-ring cell adenocarcinoma cells in endoscopic gastric biopsies.     

Fine-needle aspiration cytology diagnosis of colloid nodule versus follicular variant of papillary carcinoma of the thyroid

The cytologic differential diagnosis of colloid nodule (CN) and the follicular variant of papillary carcinoma (FVPC) is difficult with common morphologic features. To assess the utility of 18 cytologic morphometric parameters in the diagnosis of these thyroid lesions we evaluated 31 FNA samples that had histologic confirmation of the diagnoses. These 31 cases included 15 cases of CN, 8 cases of FVPC, and 8 cases of the usual variant of papillary carcinoma (UVPC) for reference values. For the morphometric analysis we used an Optimas 4.0 image analysis system. Comparing the CN group with the UVPC group revealed that eight of the parameters had statistically significant differences. The UVPC specimens were more cellular, less cohesive, had presence of papillary cellular groups more frequently, larger nuclei (UVPC: 109.33 ± 30.19 μm²; CN: 66.81 ± 15.02 μm2), higher nuclear to cytoplasmic (N/C) ratio, larger nucleoli, and present nuclear grooves and nuclear pseudoinclusions more frequently. The FVPC group differed from the CN group only in three parameters which included larger nuclei (98.49 ± 18.24 μm²), higher N/C ratio, and a more frequent presence of nuclear pseudoinclusions. When we compared these two variants of papillary carcinoma, we found that the UVPC specimens had less cellular cohesion, less preservation of the architectural polarity and a more frequent presence of papillary cellular groups than the FVPC. The FVPC can be differentiated from CN based on nuclear changes, which included a larger size, higher N/C ratio, and presence of pseudoinclusions. The absence of cellular cohesion and polarity combined with the presence of papillary groups are useful in separating the UVPC from the FVPC. A cutoff of 75 μm² should be used in separating benign from malignant nuclei. Diagn. Cytopathol. 1998;18:87–90. © 1998 Wiley-Liss, Inc.     

c-sis反义寡核苷酸对肺血管平滑肌细胞增殖和细胞周期的影响

目的:探讨从血小板源生长因子-B链(PDGF-B)的基因水平阻断对肺血管平滑肌细胞增殖的影响。方法:给肺动脉平滑肌细胞(VSMC)分别施加不同剂量的c-sis癌基因反义寡核苷酸(ASON), 通过四甲基偶氮唑蓝(MTT)法观察细胞不同时相的增殖曲线;采用流式细胞术观察不同干预条件下VSMC细胞周期的变化。结果:中、大浓度ASON对细胞增殖有明显抑制作用。在中浓度的ASON作用下, 加入PDGF-BB能促进VSMC的增殖。ASON干预前后, VSMC增殖周期中各期细胞构成发生显著的变化, 以G₀＋G₁期细胞增多、S+G₂＋M期细胞减少为特征。各组的G₀＋G₁期细胞均显著多于对照组(P＜0.05)。小剂量与中剂量、中剂量与大剂量间G₀＋G₁期细胞有显著差异(P＜0.05)。结论:中、大剂量ASON能明显抑制肺VSMC的增殖, 可使G₀＋G₁期细胞数明显增多, 且与ASON有剂量依赖关系。     

Computerized classification of nuclear profiles in non-Hodgkin's lymphomas

The authors present experimental technics for the diagnosis of non-Hodgkin's lymphomas, based on instrumental classification of nuclear profiles using a video-based system for computerized interactive morphometry (CIM). In their system, the real time video image of a specimen is superimposed to a graphics overlay generated by a computer, consisting of a test area with four visual markers for random sampling of cells and a menu with several options to send direct commands to the system. Using a touch-sensitive screen mounted on a video monitor as an interactive peripheral, a trained observer traces 100 randomly selected lymphoid cells, counts mitoses in 25 microscopic fields, and categorizes the lesion as diffuse or nodular. Each cell is instrumentally classified into either small cell noncleaved, small cell cleaved, or large, based on the length of their nuclear profiles, their enclosed nuclear area, and a circularity factor. Thereafter the computer provides a "diagnosis," based on hierarchic analysis of the data. The morphometric data are also interpreted by alternate statistical methods of discriminatory classificatory analysis that provide a diagnosis and a probability statement derived from matching unknown cases with a data base. Forty-two lymphoid lesions have been categorized with the CIM system. Studies of interobserver and intraobserver variations in data collection are discussed. The potential advantages of CIM for the objective classification of non-Hodgkin's lymphomas are discussed.     

Nuclear profiles of cancer cells reveal the metastatic potential of gastric cancer

Nuclear profiles have been reported as useful prognostic predictors in various cancers. Data from computerized morphometry are objective and can be quickly derived using conventional microscopic analysis, but image analysis of nuclear features has only rarely been applied to investigations of gastric cancer. The aim of this study was to evaluate the correlation between one of these morphological nuclear features and the clinicopathological parameters in patients with gastric cancer. The morphometric nuclear feature (nuclear area) was analysed in 400 patients with gastric cancer. In each case, 300 cancer nuclei on routine haematoxylin and eosin-stained slides were analysed through the use of a computer-assisted image analysis system which traced the nuclear profiles (magnificationx400) on a computer monitor. The morphometric data were compared with the patients' clinicopathological status and survival rate. The mean nuclear area (NA) of cancer cells from 400 cases of gastric cancer was 47.2 microm(2). The NAs of cancer cells from tumours with microvessel invasion (lymphatic or venous invasion), lymph node metastasis or hepatic metastasis at the time of operation were significantly larger than those of cancer cells from tumours without such invasion or metastases. Cytokeratin (CK) immunostaining was performed on 2577 lymph nodes from 91 patients with advanced gastric cancer (pT3, pN0, pM0, stage II) to detect micrometastases. CK-positive lymph nodes were detected in 350 of 2577 lymph nodes (13. 6%) and in 62 of 91 patients (68.1%). The mean NA of cancer cells from 62 tumours with micrometastases (44 microm(2)) was larger than that of cancer cells from 29 tumours without micrometastases (38.8 microm(2), p=0.043), and a significant positive correlation was detected between the NAs of cancer cells from 91 tumours and the number of micrometastatic lymph nodes of 91 patients (rho=0.278, p=0. 008). Cancer cells with large NA correlated strongly with haematogenous and lymph node recurrence or relapse after gastrectomy and the NA of cancer cells was identified as an independent prognostic factor in gastric cancer. Nuclear morphometry is an objective, reproducible, and technically uncomplicated procedure. The NA of cancer cells correlates closely with the metastatic potential of gastric cancer. Nuclear morphometry may therefore be useful for the selection of patients who are at risk of haematogenous or lymph node metastatic recurrence after surgery.     

Clinical significance of the histomorphometric evaluation of diabetic microangiopathy in the oral mucosa

Several structural parameters of the capillary vessels were measured in the oral mucosa of patients with diabetes mellitus of type 1 (D.1) and of type 2 (D.2), and of control cases (C), by means of an image analyser in histological sections of routinely processed biopsies. The studied parameters included: a) capillary wall thickness; b) capillary diameter; c) the ratio of capillary wall thickness and diameter; d) capillary wall area; e) capillary area; f) the ratio of capillary wall area and capillary area; g) density of capillary vessels in the lamina propria; h) density of endothelial cells; i) endothelial nuclear area. Clinical and laboratory parameters were also evaluated (duration of the disease, systolic and diastolic blood pressure levels, glycemia, glycosylated haemoglobin, glycosylated albumin, fructosamines, apolipoproteins A1 and B), in order to assess whether a relationship exists with the morphometric parameters studied. Statistically significant differences, at the level of p less than 0.05, were found in the following morphometric parameters between controls and each group of diabetic patients: mean and standard deviation of capillary wall thickness, mean capillary wall area, mean ratio of the capillary wall area and capillary area. A reduction in the capillary density, i.e. the number of capillary vessels per mm2 of lamina propria, was also observed in diabetic patients with respect to the control group, although it was not statistically significant (C vs. D.1: p less than 0.21; C vs. D.2: p less than 0.10).(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantitative analysis of non-Hodgkin's lymphoma.

A preliminary attempt has been made to characterise a small series of non-Hodgkin''s lymphomas (NHL) by morphometric means using the Quantimet 720 Kontron MOP/AMO3 image analysis systems. In most cases it was found that the distribution of nuclear area and correlation between mean nuclear area and frequency per unit field, corresponded closely with tumour classification determined by light microscopy. These results suggest that it may be possible to devise an objective and reproducible grading system for NHL using quantitative morphometric techniques.     

An ultrastructural morphometric and immunophenotypic evaluation of Burkitt's and Burkitt's-like lymphomas

Six cases of Burkitt's lymphoma (BL) and 12 cases of Burkitt's-like lymphoma (BLL) were classified by using strict histologic and cytologic criteria. These cases were processed for electron microscopy and analyzed by using computerized image analysis techniques. Form factor (4 pi A/P2) was used to measure nuclear contour irregularity. The mean of the standard deviation (SD) of nuclear area and form factor was used to assess pleomorphism. Overall, there were 8 similarities and 10 statistically significant dissimilarities out of 18 parameters analyzed. The similarities (p greater than 0.05) between the BL and BLL groups included the means of form factor, nuclear area/cytoplasmic area, SD of nuclear area/cytoplasmic area, number of nuclear pockets/100 nuclei, percentage of cells with nuclear pockets, number of lipid droplets/micron 2 of cytoplasm, percentage of cells with lipid droplets and number of mitochondria/micron 2 of cytoplasm. The dissimilarities (p less than 0.05) included the means of nuclear perimeter, SD of nuclear perimeter, nuclear area, SD of nuclear area, cellular area, SD of cellular area, cytoplasmic area, SD of cytoplasmic area, SD of form factor, and nucleolar frequency. Multiparameter analysis clearly separated these 18 patients into two distinct groups and confirms that the subtleties used in the histologic classification of these lymphoma subtypes are meaningful. Sixteen cases of BL and BLL were snap-frozen in isopentane and analyzed by using 16 lymphoid surface markers. All of the immunoglobulin-positive BL were of the mu isotype, whereas the BLL cases were divided between mu (6 cases) and gamma expression (4 cases). All 4 of the BL evaluated manifested CALLA expression, whereas 3 of the 11 BLL evaluated coexpressed CALLA. One BL case was of a pre-pre-B phenotype and one BLL case was of pre-B phenotype. The BL and BLL were compared to 49 cases of SIg (+) large cell lymphomas. The high incidence of coexpression of lambda, CALLA, and Ki-67 in BL and BLL separates these lymphomas, as a group, from the large cell lymphomas. We have also determined from this study that the separation of patients into distinct BL and BLL subtypes is clinically relevant. The BL group were all children (median of 6.5 years) compared with the BLL group who were all adults (median of 63 years). The complete remission rate was higher in the BL (67%) than in the BLL group (25%).4off     

A morphometric approach to the grading of chondroid tumours on fine-needle smears

A morphometric study was performed on 600 nuclei from 6 benign chondroid tumours (BCL) (3 enchondromas and 3 exostoses), 600 nuclei from 6 well-differentiated chondrosarcomas (WDC) and 200 nuclei from moderately-differentiated chondrosarcomas (MDC) obtained by fine needle aspiration biopsy. The parameters considered were nuclear area, perimeter and maximum diameter. Moving across the three groups of lesions the nuclei appeared progressively larger. A statistical analysis was performed on the three groups. Significant differences between the means of all the parameters considered were observed. The authors suggest that morphometric analysis may be a possible step in the cytologic diagnosis and grading of chondroid tumours.     

Image Analysis in Non-Hodgkin's Lymphoma

Abstract

Morphometry allows an objective method of assessing various cell features that are relevant in the subtyping of non Hodgkin's lymphoma (NHL). A wide variety of morphometric parameters that quantify features such as nuclear size and shape can be obtained with fully automated or interactive computerized image analysis. Most of these parameters rely on simple measures of area and perimeter, whereas others, especially those describing shape, use complicated mathematical formulas. Certain parameters have been found to be useful in specific situations; for example, the identification of cells of mycosis fungoides and Sézary syndrome is aided by the nuclear contor index. The application of morphometry in the subtyping of NHL is reviewed. Examples of cytologic preparations are given to illustrate theuse of nuclear size parameters to distinguish small, mixed, and large cell subtypes of follicular NHL. Large cell lymphoma and irnrnunoblastic sarcoma are categorized by nuclear area and shape, nucleolar area, and the standard deviation of nuclear-cytoplasmic ratio. Morphometric analysis is a useful method to subtype difficult cases of malignant lymphoma, particularly in identifying the controversial category of mixed cell lymphoma. (The J Histotechnol 15:263-276, 1992)     

Image analysis of lymphoid cell differentiation in rat thymus throughout development.

In order to identify subtle changes in cell morphology and nuclear pattern modification during thymus ontogenesis, cell image analysis using System for Analytical Microscopy in Biological Applications (SAMBA 200) was applied in 9 stages of rat thymus development. The morphometric and chromatin parameters made it possible not only to identify automatically between the two main cell populations in the thymus gland (lymphoid and epithelial cells), but also to classify automatically 5 lymphoid sub-populations (lymphoid stem cells, lymphoblasts, large lymphocytes, medium lymphocytes and small lymphocytes). The evaluation of the 18 parameters during the lymphoid cell differentiation was studied in detail. The nuclear texture parameters made it possible to discriminate, in each cell subpopulation, 4 phases of cell cycle (G0, G1, S-phase, and cells in G2). Evaluation of the nuclear parameters of the cell cycle in each lymphoid sub-population was studied in this investigation. The results illustrate the high majority of the lymphoid stem cells at the 14-day-old embryo stage while in the 20-day-old embryo the small lymphocytes become the main part of the whole lymphoid population. From the continuously renewed modification of lymphoid nuclear image analysis we discuss the origin of thymus lymphocytes. Lymphoid cells can be distinguished into different functional states and the striking morphological changes appearing during cell differentiation are related with drastic structural changes occurring in chromatin pattern from undifferentiated lymphoid stem cells to small lymphocytes. Terminal cell differentiation is associated with inhibited cell proliferation. The relative increase of chromatin condensation and nuclear pattern heterogeneity which reaches an extreme in small lymphocytes is accompanied by a progressive diminution of the nuclear area during the successive differentiation of the lymphoid population. Using one parameter of the nuclear texture features from the co-occurrence matrix (as LM or CON) and one parameter of the nuclear textures from the run-length section matrix (as GLD or RPC) the image analysis can discriminate between the different states of lymphoid cell differentiation.